Abstract:

Objective: To investigate the effect of curcumin - mediated exosomes of bone marrow mesenchymal stem cells (BMSCs) on proliferation and metastasis of ACC-M cells and its mechanism. Methods: After being co-cultured with 7.5 μmmol/L curcumin and BMSCs for 72 h, exosomes were isolated and identified, and co-cultured with ACC-M cells. ACC-M cells were divided into control group (normal culture), BMSC-Exo group (no curcumin-treated BMSCs exosomes co-cultured with ACC-M cells), Cur-BMSC-Exo group (curcumin-treated BMSCs exosomes co-cultured with ACC-M cells) and Cur group (7.5 μmmol/L curcumin treated cells). Western blotting was used to detect the expression of tumor susceptibility gene 101 (TSG-101), CD63, CD9 and recombinant human calnexin (CANX) proteins in exosome and the expression of E-cadherin, N-cadherin, vimentin, transforming growth factor-β1 (TGF-β1) and p-ERK proteins in ACC-M cells after 48 h of culture. The survival rate of ACC-M cells was determined by cell counting kit-8 (CCK-8) assay. The migration and invasion of ACC-M cells were detected by transwell assay. The relative expression levels of TGF-β1 and ERK mRNA in ACC-M cells were detected by real-time quantitative polymerase chain reaction(RT-qPCR) assay.Results: Compared with control group and BMSC-Exo group, the cell viability, cell migration and invasion number, and the expression levels of N-cadherin, vimentin, TGF-β1 and ERK were significantly decreased in Cur-BMSC-Exo and Cur groups (P<0.05), while E-cadherin protein level was significantly increased (P<0.05); compared with the Cur-BMSC-Exo group, the cell viability, cell migration and invasion numbers, and the expression levels of N-cadherin, vimentin, TGF-β1 and ERK were significantly increased in the Cur group (P<0.05), while E-cadherin protein level was significantly decreased (P<0.05). Conclusion: BMSCs exosomes can act as curcumin carrier to inhibit proliferation and metastasis of ACC-M cells and regulate TGF-β1/ERK signaling pathway.

Key words: curcumin, bone marrow mesenchymal stem cells, exosome, ACC-M, proliferation

摘要：

目的：探讨姜黄素介导的骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）外泌体对ACC-M细胞增殖和转移的影响及其作用机制。方法：将7.5 µmmol/L的姜黄素与人BMSCs共培养72 h后，分离外泌体并进行鉴定，将外泌体与ACC-M细胞进行共培养，并将ACC-M细胞分为control组（正常培养）、BMSC-Exo组（未经姜黄素处理的BMSCs外泌体与ACC-M细胞共培养）、Cur-BMSC-Exo组（经姜黄素处理的BMSCs外泌体与ACC-M细胞共培养）及Cur组（经7.5 µmmol/L的姜黄素处理细胞）；培养48 h后使用蛋白质印迹法（Western blotting）实验检测外泌体肿瘤易感基因101（tumor susceptibility gene 101，TSG-101）、CD63、CD9和重组人钙连蛋白(calnexin，CANX)及ACC-M细胞上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）和转化生长因子β1（transforming growth factor-β1，TGF-β1)、细胞外信号调节激酶（extracellular signal-regulated kinase，ERK)的蛋白表达情况；使用细胞计数试剂盒-8（cell counting kit-8，CCK8）实验检测ACC-M细胞存活率；使用transwell实验检测ACC-M细胞迁移和侵袭情况；使用实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR）检测ACC-M细胞TGF-β1和ERK mRNA相对表达量。结果：与control组和BMSC-Exo组比较，Cur-BMSC-Exo组和Cur组的细胞活力、细胞迁移和侵袭数量及N-cadherin、vimentin、TGF-β1和ERK 表达水平均显著降低（P<0.05），而E-cadherin蛋白水平显著增加（P<0.05）；与Cur-BMSC-Exo组比较，Cur组细胞活力、细胞迁移和侵袭数量及N-cadherin、vimentin、TGF-β1和ERK 表达水平均显著增加（P<0.05），而E-cadherin蛋白水平显著降低（P<0.05）。结论：BMSCs分泌的外泌体可作为姜黄素的载体，抑制ACC-M细胞的增殖和转移并调控TGF-β1/ERK信号通路。

关键词: 姜黄素, 骨髓间充质干细胞, 外泌体, ACC-M, 增殖

CLC Number: 

• R782